Journal of Pharmaceutical Research

Open Access Open Access  Restricted Access Subscription Access

Analytical Method Development and Validation of Metformin, Losartan and Glimepiride in Bulk and Combined Tablet Dosage form by Gradient RP-HPLC


Affiliations
1 Department of Pharmaceutical Analysis, University College of Pharmaceutical Science, Acharya Nagarjuna University , Nagarjuna Nagar, Guntur – 522510, AP, India
 

Purpose: A simple, sensitive, linear, precise, and accurate method by gradient reversed-phase-high performance liquid chromatography for the simultaneous estimation of metformin (MET), losartan (LOS) and glimepiride (GLI) in bulk and in their combined tablet dosage form was developed and validated.

Methodology: The separation of the three drugs was based on the use of Luna c18 (250 ~ 4.6 mm, i.e. 5 ƒÊm) column in a gradient mode. Mobile phase consisted of Methanol (solvent A) and 0.1% Orthophosphoric acid [OPA] (solvent B) was set with gradient programming for 18 min and was delivered at 1 ml/min flow rate and effluents are achieved with variable wavelength: Photodiode array detector at 284 nm. The retention times of MET, LOS and GLI were found to be 3.11, 7.12 and 13.52mins respectively. The percentage assay of MET, LOS and GLI was found to be 100.5%, 100.5 and 100.4%, respectively. Calibration curves were linear for MET, LOS and GLI at concentration ranges of 30- 450 ng/ml, and 15-225ng/ml and 1-18ng/ml with the regression coefficient of 0.999 for all the three drugs and precise with (% RSD <2). The drug was subjected to various stress conditions of acid and base hydrolysis, oxidation, photolysis, thermal degradation and condition.

Findings: Considerable degradation was found under all stress conditions and the degradation products were well resolved from Metformin (MET),Llosartan (LOS) and Glimepiride (GLI) in the proposed gradient RP-HPLC method.

Conclusion: The method was validated by determining its linearity, accuracy, precision, system suitability and can be employed for routine quality control analysis.


Keywords

RP-HPLC, Validation, Stability Studies, ICH Guidelines.
User
Notifications
Font Size


  • Available from: http://www.drugbank.ca/drugs/DB00883 Metformin HCL.

  • Available from: http://en.wikipedia.org/ Losartan.

  • Available from: http://www.drugbank.ca/drugs/DB01275 Glimepiride.

  • Kral K, Kainz G. Simultaneous determination of Metformin, losartan, glimepiride in pharmaceutical preparations by HPLC with electrochemical detection. Fresenius' Z. Anal. Chem. 1983; 316: 497-500.

  • Torok I, Paal T, Koszegi-Szalai H, Keseru P. High-performance liquid chromatographic assay and content uniformity determination of metformin and losartan in tablets. Acta Pharm. Hung. 1985; 55: 154-62.

  • Neelima K, Rajendra Prasad Y. Analytical method development and validation for simultaneous estimation of metformin, losartan, glimepiride in bulk and tablet formulation by RP-HPLC. International Journal of Pharmaceutical Sciences and Research, 2014; 5(4): 1290-1294.

  • Molles RJ, Garceau Y. Quantitation of metformin, losartan, glimepiride in pharmaceutical dosage forms by high-performance liquid chromatography. J Chromatogr A. 1985; 347:414-418.

  • Wong JK, Joyce TH, Morrow DH. Determination of Metformin in human plasma by high-performance liquid chromatography with electrochemical detection. J Chromatogr A. 1987; 385:261-266.

  • Smith KM, Johnson RN, Kho BT. Determination of glimepiride in tablets by gas chromatography. J Chromatogr A. 1977; 137(2):431-437.

  • Manes J, Mari J, Garcia R, Font G. Liquid chromatographic determination of metformin & losartan in human plasma with 2-hydroxy-1naphthaldehyde pre-column derivatization. J Pharm Biomed Anal. 1990; 8(12):795-798.

  • Wong JK, Joyce TH, Morrow DH. Determination of glimepiride in human plasma by high performance liquid chromatography with electrochemical detection. J Chromatogr A. 1987; 385:261-266. http://dx.doi.org/10.1016/S0021-9673 (01)94638-2.

  • Olsen C S, Scroggins H S. High-performance liquid chromatographic determination of the nitrate esters metformin, losartan, glimepiride in various tablet forms. J. Pharm. Sciences.1983; 73: 1303-4.

  • Woo D, Yen J.K.C, Sofronas P. Quantitative analysis of metformin, losartan, glimepiride by infrared spectrometry. Anal. Chem.1973 45: 2144-45.

  • Stewart JT, Parks EH. Colorimetric determination of Glimepiride with 9methoxyacridine. Int J Pharm. 1983;17(2-3):161-1.

  • Stewart JT, Parks EH. Colorimetric determination of Metformin with 9methoxyacridine. Int J Pharm. 1983; 17(2-3):161-166.

  • Smith KM, Johnson RN, Kho BT. Determination of Metformin, losartan tablets by Gas chromatography. J Chromatogr A. 1977; 137(2):431-437.

  • Degen PH. Determination of unchanged Metformin, Losartan, Glimepiride in plasma by Gas-liquid chromatography using nitrogenspecific detection. J Chromatogr A. 1979; 176(3):375-380.

  • Klaus D, Haegele, Harold B, Skrdlant, Norman W, Robie, David Lalka, John L, McNay Jr. Determination of metformin and its metabolites by gas chromatography-mass spectrometry. Journal of Chromatography A. 1976; 126 (3):517.


Abstract Views: 196

PDF Views: 63




  • Analytical Method Development and Validation of Metformin, Losartan and Glimepiride in Bulk and Combined Tablet Dosage form by Gradient RP-HPLC

Abstract Views: 196  |  PDF Views: 63

Authors

Mastanamma Shaik
Department of Pharmaceutical Analysis, University College of Pharmaceutical Science, Acharya Nagarjuna University , Nagarjuna Nagar, Guntur – 522510, AP, India
Maharshi Viswanadhapalli
Department of Pharmaceutical Analysis, University College of Pharmaceutical Science, Acharya Nagarjuna University , Nagarjuna Nagar, Guntur – 522510, AP, India
Saidulu Pasupuleti
Department of Pharmaceutical Analysis, University College of Pharmaceutical Science, Acharya Nagarjuna University , Nagarjuna Nagar, Guntur – 522510, AP, India
Reehana Shaik
Department of Pharmaceutical Analysis, University College of Pharmaceutical Science, Acharya Nagarjuna University , Nagarjuna Nagar, Guntur – 522510, AP, India

Abstract


Purpose: A simple, sensitive, linear, precise, and accurate method by gradient reversed-phase-high performance liquid chromatography for the simultaneous estimation of metformin (MET), losartan (LOS) and glimepiride (GLI) in bulk and in their combined tablet dosage form was developed and validated.

Methodology: The separation of the three drugs was based on the use of Luna c18 (250 ~ 4.6 mm, i.e. 5 ƒÊm) column in a gradient mode. Mobile phase consisted of Methanol (solvent A) and 0.1% Orthophosphoric acid [OPA] (solvent B) was set with gradient programming for 18 min and was delivered at 1 ml/min flow rate and effluents are achieved with variable wavelength: Photodiode array detector at 284 nm. The retention times of MET, LOS and GLI were found to be 3.11, 7.12 and 13.52mins respectively. The percentage assay of MET, LOS and GLI was found to be 100.5%, 100.5 and 100.4%, respectively. Calibration curves were linear for MET, LOS and GLI at concentration ranges of 30- 450 ng/ml, and 15-225ng/ml and 1-18ng/ml with the regression coefficient of 0.999 for all the three drugs and precise with (% RSD <2). The drug was subjected to various stress conditions of acid and base hydrolysis, oxidation, photolysis, thermal degradation and condition.

Findings: Considerable degradation was found under all stress conditions and the degradation products were well resolved from Metformin (MET),Llosartan (LOS) and Glimepiride (GLI) in the proposed gradient RP-HPLC method.

Conclusion: The method was validated by determining its linearity, accuracy, precision, system suitability and can be employed for routine quality control analysis.


Keywords


RP-HPLC, Validation, Stability Studies, ICH Guidelines.

References